U.S. patent application number 11/493985 was filed with the patent office on 2007-02-08 for systems, methods, and compositions for modifying the metabolism of plants and of euucaryotic microbes.
Invention is credited to M. Karen Newell.
Application Number | 20070032383 11/493985 |
Document ID | / |
Family ID | 37718325 |
Filed Date | 2007-02-08 |
United States Patent
Application |
20070032383 |
Kind Code |
A1 |
Newell; M. Karen |
February 8, 2007 |
Systems, methods, and compositions for modifying the metabolism of
plants and of euucaryotic microbes
Abstract
Systems, methods, and compositions for modifying the metabolsim
of plants and of eucaryotic microbes, and for culturing them for
the production of lipids with high concentrations of omega-3 highly
unsaturated fatty acids (HUFA) suitable for human and animal
consumption as food additives or for use in pharmaceutical and
industrial products. The metabolic modifier is added to a
fermentation or growth medium, and is selected from the group
consisting essentially of a fatty acid metabolism inhibitor,
glycolysis inhibitor, a bifunctional compound that links a fatty
acid metabolism inhibitor to a glycolysis inhibitor, glucose,
insulin, and a plant growth factor.
Inventors: |
Newell; M. Karen; (Colorado
Springs, CO) |
Correspondence
Address: |
BERLINER & ASSOCIATES
555 WEST FIFTH STREET
31ST STREET
LOS ANGELES
CA
90013
US
|
Family ID: |
37718325 |
Appl. No.: |
11/493985 |
Filed: |
July 27, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60703232 |
Jul 27, 2005 |
|
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Current U.S.
Class: |
504/291 ;
514/475 |
Current CPC
Class: |
A01N 37/44 20130101;
A01N 43/08 20130101; A01N 43/40 20130101; Y02P 60/20 20151101; A01N
43/12 20130101; A01N 37/50 20130101; A01N 43/60 20130101; Y02P
60/246 20151101; A01N 53/00 20130101; A01N 33/10 20130101; A01N
37/36 20130101; A01N 37/06 20130101 |
Class at
Publication: |
504/291 ;
514/475 |
International
Class: |
A01N 43/02 20060101
A01N043/02; A01N 43/20 20060101 A01N043/20 |
Claims
1. A method for modifying the metabolsim of plants or eucaryotic
microbes, and for culturing them for the production of lipids with
high concentrations of omega-3 highly unsaturated fatty acids,
comprising feeding the plant or eukaryotic microbe a metabolic
modifier selected from the group consisting essentially of a fatty
acid metabolism inhibitor, glycolysis inhibitor, a bifunctional
compound that links a fatty acid metabolism inhibitor to a
glycolysis inhibitor, glucose, insulin, a plant growth factor, and
inhibitors of gluconeogenesis and CO2 fixation in plants that
indirectly promote storage of fat.
2. The method of claim 1 in which the metabolic modifier is added
to a fermentation or growth medium
3. The method of claim 1 in which the bifunctional compound links a
fatty acid inhibitor to an inhibitor that is, or is derived from,
or has the functionality of, hypoglycin A.
4. The method of claim 1 in which the fatty acid metabolism
inhibitor has the functionality of an oxirane carboxylic acid
compound.
5. The method of claim 1 in which the glycolysis inhibitor has the
functionality of 2-deoxy-D-glucose.
6. The method of claim 1 in which the bifunctional compound links a
moiety having the functionality of etomoxir to a moiety having the
functionality of hypoglycin A.
7. The method of claim 1 in which the bifunctional compound links a
moiety having the functionality of etomoxir to a moiety having the
functionality of 2-deoxy -D-glucose.
8. The method of claim 1 in which the eucaryotic microbes are
selected from the group consisting of algae, fungi, protists, and
mixtures thereof, wherein the microorganisms are capable of
producing polyenoic fatty acids or other lipids which requires
molecular oxygen for their synthesis.
9. The method of claim 8 in which the eukaryotic microorganisms are
capable of producing lipids at a fermentation medium oxygen level
of about less than 3% of saturation.
10. The method of claim 1 in which the fatty acid metabolism
inhibitor is selected from the group consisting of: (a) carboxylic
acids having the structure: ##STR48## where R comprises an organic
moiety and may include at two or more nitrogen atoms, or an
aromatic moiety; (b) compounds having the structure: ##STR49##
where each of R.sup.1 and R.sup.2 independently comprises organic
moiety in which either or both of R.sup.1 and R.sup.2 may
independently be an alkyl, may include an aromatic moiety, or may
have the structure: ##STR50## where R.sup.3 comprises an organic
moiety and Ar.sup.1 comprises an aromatic moiety. R.sup.3 may be an
alkyl, such as a straight-chain alkyl.; (c) compounds having the
structure: ##STR51## where each of R.sup.10, R.sup.11, R.sup.12,
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 independently comprises
hydrogen, a halogen, or an organic moiety, or R.sup.10 and R.sup.11
together may define an organic moiety; and (d) compounds having the
structure: ##STR52## wherein: R.sub.1 represents a hydrogen atom, a
halogen atom, a 1-4C alkyl group, a 1-4C alkoxy group, a nitro
group or a trifluoromethyl group; R.sub.2 has one of the meanings
of R.sub.1; R.sub.3 represents a hydrogen atom or a 1-4C alkyl
group; Y represents the grouping --O--(CH.sub.2).sub.m--; m is 0 or
a whole number from 1 to 4; and n is a whole number from 2 to 8
wherein the sum of m and n is a whole number from 2 to 8.
11. The method of claim 1 in which the glycolysis inhibitor has the
structure: ##STR53## wherein: X represents an O or S atom; R.sub.1
represents a hydrogen atom or a halogen atom; R.sub.2 represents a
hydroxyl group, a halogen atom, a thiol group, or CO--R.sub.6; and
R.sub.3, R.sub.4, and R.sub.5 each represent a hydroxyl group, a
halogen atom, or CO-- R.sub.6 wherein R.sub.6 represents an alkyl
group of from 1 to 20 carbon atoms, and wherein at least two of
R.sub.3, R.sub.4, and R.sub.5 are hydroxyl groups.
12. The method of claim 1 in which the glycolysis inhibitor is
hypoglycin A. The method of claim 1 in which the bifunctional
compound is selected from the group consisting of: (a) compounds
having the structure: ##STR54## where R.sub.1 represents a hydroxyl
group, a halogen atom, a thiol group, or CO--R.sub.9 where R.sub.9
represents an alkyl group of from 1 to 20 carbon atoms, R.sub.4
represents a hydrogen atom or a halogen atom, R.sub.2, R.sub.3 and
R.sub.5 each represent a hydroxyl group, a halogen atom, or
CO--R.sub.9 and where at least two of R.sub.2, R.sub.3 and R.sub.5
are hydroxyl groups, R.sub.6 and R.sub.7 each represent a hydrogen
atom, a halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon
atom alkoxy group, a nitro group or a trifluoromethyl group,
R.sub.8 represents a hydrogen atom or a 1-4 carbon atom alkyl
group, X represents O or S, Y represents(CH.sub.2).sub.k where k is
from 2 to 8, or the grouping --O--(CH.sub.2).sub.m--, m is 0 or a
whole number from 1 to 4, n is a whole number from 2 to 8 wherein
the sum of m and n is a whole number from 2 to 8, and Z represents
O, S or the grouping (CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (b) compounds having the structure:
##STR55## where R.sub.4 represents a hydrogen atom or a halogen
atom, R.sub.2, R.sub.3 and R.sub.5 each represent a hydroxyl group,
a halogen atom, or CO--R.sub.9 where R.sub.9 represents an alkyl
group of from 1 to 20 carbon atoms, and where at least two of
R.sub.2, R.sub.3 and R.sub.5 are hydroxyl groups, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, X
represents O or S, and Y represents(CH.sub.2).sub.k where k is from
2 to 8, or the grouping --O --(CH.sub.2).sub.m--, m is 0 or a whole
number from 1 to 4, n is a whole number from 2 to 8 wherein the sum
of m and n is a whole number from 2 to 8; (c) compounds having the
structure: ##STR56## where R.sub.1 represents a hydroxyl group, a
halogen atom, a thiol group, or CO--R.sub.9 where R.sub.9
represents an alkyl group of from 1 to 20 carbon atoms, R.sub.4
represents a hydrogen atom or a halogen atom, R.sub.2, R.sub.3 and
R.sub.5 each represent a hydroxyl group, a halogen atom, or
CO--R.sub.9 and where at least two of R.sub.2, R.sub.3 and R.sub.5
are hydroxyl groups, R.sub.6 and R.sub.7 each represent a hydrogen
atom, a halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon
atom alkoxy group, a nitro group or a trifluoromethyl group,
R.sub.8 represents a hydrogen atom or a 1- 4 carbon atom alkyl
group, X represents O or S, and Y represents(CH.sub.2).sub.k where
k is from 2 to 8, or the grouping --O--(CH.sub.2).sub.m--, m is 0
or a whole number from 1 to 4, n is a whole number from 2 to 8
wherein the sum of m and n is a whole number from 2 to 8; (d)
compounds having the structure: ##STR57## where R.sub.1 represents
a hydroxyl group, a halogen atom, a thiol group, or CO--R.sub.9
where R.sub.9 represents an alkyl group of from 1 to 20 carbon
atoms, R.sub.4 represents a hydrogen atom or a halogen atom,
R.sub.2, R.sub.3 and R.sub.5 each represent a hydroxyl group, a
halogen atom, or CO--R.sub.9 and where at least two of R.sub.2,
R.sub.3 and R.sub.5 are hydroxyl groups, R.sub.6 and R.sub.7 each
represent a hydrogen atom, a halogen atom, a 1-4 carbon atom alkyl
group, a 1-4 carbon atom alkoxy group, a nitro group or a
trifluoromethyl group, R.sub.8 represents a hydrogen atom or a 1-4
carbon atom alkyl group, X represents O or S, Y represents
(CH.sub.2).sub.k where k is from 2 to 8, or the grouping
--O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1 to 4, n is
a whole number from 2 to 8 wherein the sum of m and n is a whole
number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (e) compounds having the structure:
##STR58## where R.sub.1 represents a hydroxyl group, a halogen
atom, a thiol group, or CO--R.sub.9 where R.sub.9 represents an
alkyl group of from 1 to 20 carbon atoms, R.sub.4 represents a
hydrogen atom or a halogen atom, R.sub.2, R.sub.3 and R.sub.5 each
represent a hydroxyl group, a halogen atom, or CO--R.sub.9 and
where at least two of R.sub.2, R.sub.3 and R.sub.5 are hydroxyl
groups, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, X
represents O or S, and Y represents (CH.sub.2).sub.k where k is
from 2 to 8, or the grouping --O--(CH.sub.2).sub.m--, where m is 0
or a whole number from 1 to 4, n is a whole number from 2 to 8
wherein the sum of m and n is a whole number from 2 to 8; (f)
compounds having the structure: ##STR59## where R.sub.1, R.sub.2,
R.sub.3, R4, and R.sub.5 represents a hydroxyl group, a halogen
atom, a thiol group, or CO--R.sub.9 where R.sub.9 represents an
alkyl group of from 1 to 20 carbon atoms, R.sub.6 and R.sub.7 each
represent a hydrogen atom, a halogen atom, a 1-4 carbon atom alkyl
group, a 1-4 carbon atom alkoxy group, a nitro group or a
trifluoromethyl group, R.sub.8 represents a hydrogen atom or a 1-4
carbon atom alkyl group, Y represents (CH.sub.2).sub.k where k is
from 2 to 8, 7or the grouping --O--(CH.sub.2).sub.m--, m is 0 or a
whole number from 1 to 4, n is a whole number from 2 to 8 wherein
the sum of m and n is a whole number from 2 to 8, and Z represents
O, S or the grouping (CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (g) compounds having the structure:
##STR60## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (h) compounds having the structure:
##STR61## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (i) compounds having the structure:
##STR62## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (j) compounds having the structure:
##STR63## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (k) compounds having the structure:
##STR64## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (I) compounds having the structure:
##STR65## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (m) compounds having the structure:
##STR66## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (n) compounds having the structure:
##STR67## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (o) compounds having the structure:
##STR68## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; (p) compounds having the structure:
##STR69## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R.sub.8
represents a hydrogen atom or a 1-4 carbon atom alkyl group, Y
represents (CH.sub.2).sub.k where k is from 2 to 8, 7or the
grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1
to 4, n is a whole number from 2 to 8 wherein the sum of m and n is
a whole number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4; and (q) compounds having the structure:
##STR70## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen atom, a
halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon atom
alkoxy group, a nitro group or a trifluoromethyl group, R
.sub.8 represents a hydrogen atom or a 1- 4 carbon atom alkyl
group, Y represents (CH.sub.2).sub.k where k is from 2 to 8, 7or
the grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from
1 to 4, n is a whole number from 2 to 8 wherein the sum of m and n
is a whole number from 2 to 8, and Z represents O, S or the
grouping (CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of Provisional Patent
Application No. 60/703,232, filed Jul. 27, 2005.
FIELD OF INVENTION
[0002] The field of the invention generally relates to systems,
methods, and compositions for modifying the metabolsim of plants
and of Eucaryotic microbes, and for culturing them for the
production of lipids with high concentrations of omega-3 highly
unsaturated fatty acids (HUFA) suitable for human and animal
consumption as food additives or for use in pharmaceutical and
industrial products.
BACKGROUND
[0003] Omega-3 highly unsaturated fatty acids (HUFAs) are of
significant commercial interest as important dietary compounds for
preventing arteriosclerosis and coronary heart disease, for
alleviating inflammatory conditions and for retarding the growth of
tumor cells. These beneficial effects are a result both of omega-3
HUFAs causing competitive inhibition of compounds produced from
omega-6 fatty acids, and from beneficial compounds produced
directly from the omega-3 HUFAs themselves. Omega-6 fatty acids are
the predominant HUFAs found in plants and animals. Commercially
available sources of omega-3 HUFAS are the microflora
Thraustochytrium and Schizochytrium which are discussed in detail
in U.S. Pat. No. 5,130,242 by William M. Barclay, the entire
disclosure of which is incorporated herein by reference. These
microflora have the advantages of being heterotrophic and capable
of high levels of omega-3 HUFA production.
[0004] Eukaryotic microbes (such as algae; fungi, including yeast;
and protists) have been demonstrated to be good producers of
polyenoic fatty acids in fermentors; see U.S. Patent Application
Publication Number 2003/0180898 by Richard B. Bailey et al.,
published Sep. 25, 2003 and entilted: "Enhanced production of
lipids containing polyenoic fatty acid by very high density
cultures of eukaryotic microbes in fermentors", incorporated herein
in its entirety.
[0005] There still exists a need for improved methods for
fermentation of these microflora and of production of plant
originating fatty acids.\
SUMMARY OF INVENTION
[0006] The invention generally relates to systems, methods, and
compositions for modifying the metabolsim of plants and of
eucaryotic microbes, and for culturing them for the production of
lipids with high concentrations of omega-3 highly unsaturated fatty
acids (HUFA) suitable for human and animal consumption as food
additives or for use in pharmaceutical and industrial products. The
metabolic modifier is added to a fermentation or growth medium, and
is selected from the group consisting essentially of a fatty acid
metabolism inhibitor, glycolysis inhibitor, a bifunctional compound
that links a fatty acid metabolism inhibitor to a glycolysis
inhibitor, glucose, insulin, and a plant growth factor.
[0007] In a preferred embodiment, the bifunctional compound links a
fatty acid inhibitor to an inhibitor that is, or is derived from,
or has the functionality of, hypoglycin A (also referred to as
hypoglycine A), which can serve as both a glycolysis inhibitor and
a fatty acid metabolism inhibitor. In other preferred embodiments,
the fatty acid metabolism inhibitor has the functionality of an
oxirane carboxylic acid compound. In another preferred embodiment,
the glycolysis inhibitor has the functionality of
2-deoxy-D-glucose. In specific embodiments, the invention provides
a bifunctional compound that links a moiety having the
functionality of etomoxir to a moiety having the functionality of
hypoglycin A. In other specific embodiments, the invention provides
a bifunctional compound that links a moiety having the
functionality of etomoxir to a moiety having the functionality of
2-deoxy-D-glucose.
[0008] In yet another aspect of the present invention, the
eucaryotic microbes are selected from the group consisting of
algae, fungi, protists, and mixtures thereof, wherein the
microorganisms are capable of producing polyenoic fatty acids or
other lipids which requires molecular oxygen for their synthesis. A
particularly useful microorganisms of the present invention are
eukaryotic microorganisms which are capable of producing lipids at
a fermentation medium oxygen level of about less than 3% of
saturation.
[0009] Other advantages and novel features of the present invention
will become apparent from the following detailed description of
various non-limiting embodiments of the invention when considered
in conjunction with the accompanying figures. In cases where the
present specification and a document incorporated by reference
include conflicting and/or inconsistent disclosure, the present
specification shall control.
BRIEF DESCRIPTION OF DRAWINGS
[0010] FIGS. 1A-1C illustrate certain compounds for use with the
invention;
[0011] FIG. 2 shows generic chemical structures of five embodiments
of the bifunctional compounds of this invention;
[0012] FIG. 3 shows the chemical structures of five specific
bifunctional compounds of this invention;
[0013] FIG. 4 shows generic chemical structures (I), (II), and
(III) of a first set of three embodiments of bifunctional compounds
of this invention;
[0014] FIG. 5 shows generic chemical structures (IV), (V), and (VI)
of a second set of three embodiments of bifunctional compounds of
this invention;
[0015] FIG. 6 shows generic chemical structures (VII), (VIII), and
(IX) of a third set of three embodiments of bifunctional compounds
of this invention;
[0016] FIG. 7 shows generic chemical structures (X), (XI), and
(XII) of a fourth set of three embodiments of bifunctional
compounds of this invention;
[0017] FIG. 8 shows specific chemical structures (XIII), (XIV), and
(XV) of a set of bifunctional compounds of this invention within
the generic structures, respectively (I), (II), and (III), of FIG.
1;
[0018] FIG. 9 shows specific chemical structures (XVI), (XVII), and
(XVIII) of a set of bifunctional compounds of this invention within
the generic structures, respectively (IV), (V), and (VI), of FIG.
2;
[0019] FIG. 10 shows specific chemical structures (XIX), (XX), and
(XXI) of a set of bifunctional compounds of this invention within
the generic structures, respectively (VII), (VIII), and (IX), of
FIG. 3;
[0020] FIG. 11 shows specific chemical structures (XXII), (XXIII),
and (XIV) of a set of bifunctional compounds of this invention
within the generic structures, respectively (X), (X), and (XI), of
FIG. 4; and
[0021] FIG. 12 shows specific chemical structures (XXV), (XXVI),
and (XXVII) of a set of bifunctional compounds of this invention
within the generic structures, respectively (XI), (XII), and (XII),
of FIG. 4
DETAILED DESCRIPTION
[0022] The present invention proceeds by recognizing that cells of
all types, plant as well as eucaryotic microbes, have available to
them a number of different metabolic strategies, and that by
modifying particular strategies, one can cause the plant or microbe
to generate increased yields of desired product, exemplified by
omega-6 HUFAs in plants and omega-3 HUFAs in eucaryotic microbes.
Inhibiting fatty acid metabolism enables the cell to accumulate
faty acids. Since glucose is used as a building block of fatty acid
synthesis, by inhibiting glycolysis, more glucose is available for
the production of fatty acids. Using a bifunctional compound that
links a fatty acid metabolism inhibitor to a glycolysis inhibitor
enables both fatty acid accumulation and fatty acid synthesis.
Glucose, insulin, or a plant growth factor, each have salutory
effects.
[0023] As used herein, the term "plant" is used in its broadest
sense. The term plant includes, but is not limited to, any species
of woody, ornamental or decorative, crop or cereal, fruit or
vegetable plant, and algae (e.g., Chlamydomonas reinhardtii). As
used herein, the term "cereal crop" is used in its broadest sense.
The term includes, but is not limited to, any species of grass, or
grain plant (e.g., barley, corn, oats, rice, wild rice, rye, wheat,
millet, sorghum, triticale, etc.), non-grass plants (e.g.,
buckwheat flax, legumes [soybeans] etc.), or other common plant
derived carbohydrate source, etc. As used herein, the term "crop"
or "crop plant" is used in its broadest sense. The term includes,
but is not limited to, any species of plant or algae edible by
humans or used as a feed for animals or used, or consumed by
humans, or any plant or algae used in industry or commerce.
[0024] As used herein, the term "eucaryotic microbe," which some
refer to as "eukaryotic microbe," includes fungi (including yeast),
algae, and protists. More particularly, preferred embodiments
include growing marine microorganisms, in particular algae, such as
Thraustochytrids of the order Thraustochytriales, more specifically
Thraustochytriales of the genus Thraustochytrium and
Schizochytrium, including Thraustochytriales which are disclosed in
commonly assigned U.S. Pat. Nos. 5,340,594 and 5,340,742, both
issued to Barclay, all of which are incorporated herein by
reference in their entirety. Fungi are any of numerous eukaryotic
organisms of the kingdom Fungi, which lack chlorophyll and vascular
tissue and range in form from a single cell to a body mass of
branched filamentous hyphae that often produce specialized fruiting
bodies. The kingdom includes the yeasts, molds, smuts, and
mushrooms. Algae are ny of various chiefly aquatic, eukaryotic,
photosynthetic organisms, ranging in size from single-celled forms
to the giant kelp. Algae were once considered to be plants but are
now classified separately because they lack true roots, stems,
leaves, and embryos. Protists are any of the eukaryotic,
unicellular organisms of the former kingdom Protista, which
includes protozoans, slime molds, and certain algae. The protists
now belong to the kingdom Protoctista, a new classification in most
modem taxonomic systems.
[0025] The metabolic modifier is selected from the group consisting
essentially of a fatty acid metabolism inhibitor, glycolysis
inhibitor, a bifunctional compound that links a fatty acid
metabolism inhibitor to a glycolysis inhibitor, glucose, insulin,
and a plant growth factor. Additional inhibitors may include
inhibitors of gluconeogenesis and CO2 fixation in plants that
indirectly promote storage of fat. Glucose and insulin need no
further explanation as to their nature or type.
Plant Growth Factor
[0026] The term "plant growth factor" is usually employed for plant
hormones or substances of similar effect that are administered to
plants. Classes of plant hormones (phytohormones) are auxins,
cytokinins, gibberellins, abscisic acid, ethylene, and more
recently, polyamines such as putrescine or spermidine. They are
physiological intercellular messengers that are needed to control
the complete plant lifecycle, including germination, rooting,
growth, flowering, fruit ripening, foliage and death. In addition,
plant hormones are secreted in response to environmental factors
such as abundance of nutrients, drought conditions, light,
temperature, chemical or physical stress. Hence, levels of hormones
will change over the lifespan of a plant and are dependent upon
season and environment. Plant growth factors are widely used in
industrialized agriculture to improve productivity. The application
of growth factors allows synchronization of plant development to
occur.
Fatty Acid Metabolism Inhibitors
[0027] A "fatty acid metabolism inhibitor," as used herein, is a
compound able to inhibit (e.g., prevent, or at least decrease or
inhibit the activity by an order of magnitude or more) a reaction
within the fatty acid metabolism pathway, such as an
enzyme-catalyzed reaction within the pathway. The inhibitor may
inhibit the enzyme, e.g., by binding to the enzyme or otherwise
interfering with operation of the enzyme (for example, by blocking
an active site or a docking site, altering the configuration of the
enzyme, competing with an enzyme substrate for the active site of
an enzyme, etc.), and/or by reacting with a coenzyme, cofactor,
etc. necessary for the enzyme to react with a substrate. The fatty
acid metabolism pathway is the pathway by which fatty acids are
metabolized within a cell for energy (e.g., through the synthesis
of ATP and the breakdown of fatty acids into simpler structures,
such as CO.sub.2, acyl groups, etc.).
[0028] The fatty acid metabolism pathway includes several enzymatic
reactions, which uses various enzymes such as reductases or
isomerases. Specific examples of enzymes within the fatty acid
metabolism pathway include 2,4-dienoyl-CoA reductase,
2,4-dienoyl-CoA isomerase, butyryl dehydrogenase, etc, as further
discussed below. In one embodiment, the fatty acid metabolism
inhibitor is an inhibitor able to inhibit a beta-oxidation reaction
in the fatty acid metabolism pathway. In another embodiment, the
inhibitor is an inhibitor for a fatty acid transporter (e.g., a
transporter that transports fatty acids into the cell, or from the
cytoplasm into the mitochondria for metabolism). In yet another
embodiment, the inhibitor may react or otherwise inhibit key steps
within the fatty acid metabolism pathway. In still another
embodiment, the inhibitor may be an inhibitor of fatty acids as a
source of energy in the mitochondria. For example, the inhibitor
may inhibit the breakdown of intermediates such as butyryl CoA,
glutaryl CoA, or isovaleryl CoA.
[0029] 2,4-dienoyl-CoA reductase is an enzyme within the fatty acid
metabolism pathway that catalyzes reduction reactions involved in
the metabolism of polyunsaturated fatty acids. Certain fatty acids
are substrates for 2,4-dienoyl-CoA reductases located within the
mitochondria. In some cases, fatty acids may be transported into
the mitochondria through uncoupling proteins. The uncoupling
protein may, in certain instances, increase the mitochondrial
metabolism to increase the availability of fatty acids within the
mitochondria and/or increase the throughput of beta-oxidation
within the mitochondria.
[0030] The enzyme 2,4-dienoyl-CoA isomerase is an enzyme within the
fatty acid metabolism pathway that catalyzes isomerization of
certain fatty acids. One step in the metabolism of certain
polyunsaturated fatty acids may be protective against reactive
oxygen intermediates ("ROI"). Thus, by generating substrates and
antagonists for the activity of 2,4-dienyol-CoA isomerase, the
metabolic production of reactive oxygen intermediates may be
enhanced and/or reduced. This, in turn, may affect certain disease
states, such as cancer.
[0031] Thus, it is to be understood that, as used herein, compounds
useful for inhibiting fatty acid metabolism (i.e., "fatty acid
metabolism inhibitors") are also useful for altering cellular
production of reactive oxygen; compounds described in reference to
fatty acid metabolism inhibition should also be understood herein
to be able to alter reactive oxygen production within a cell. For
example, by altering the ability of a cell to metabolize a fatty
acid, the ability of the cell to produce reactive oxygen may also
be affected, since one pathway for a cell to produce reactive
oxygen intermediates is through the metabolism of fatty acids.
Alteration of the production of reactive oxygen in a cell may be
associated with changes in the immune profile of cells, i.e., how
immune cells respond to the cell. Thus, in some cases, the
production of reactive oxygen can be affected by exposing a cell
to, or removing a cell from, a fatty acid metabolism inhibitor.
[0032] In a preferred embodiment of the invention, the fatty acid
inhibitor is an oxirane carboxylic acid compound. In accordance
with a discovery of this invention, such compounds, exemplified by
etomoxir, are able to alter cellular production of reactive oxygen.
Preferred oxirane carboxylic acid compounds have the formula:
##STR1## wherein: R.sub.1 represents a hydrogen atom, a halogen
atom, a 1-4C alkyl group, a 1-4C alkoxy group, a nitro group or a
trifluoromethyl group; R.sub.2 has one of the meanings of R.sub.1;
R.sub.3 represents a hydrogen atom or a 1-4C alkyl group; Y
represents the grouping --O--(CH.sub.2).sub.m--; m is 0 or a whole
number from 1 to 4; and n is a whole number from 2 to 8 wherein the
sum of m and n is a whole number from 2 to 8. More preferred are
oxirane carboxylic acid compounds wherein R.sub.1 is a halogen
atom, R.sub.2 is a hydrogen atom, m is 0, and n is 6, and more
particulary where R.sub.3 is an ethyl group.
[0033] It is most particularly preferred to use etomoxir, i.e.,
2-(6-(4-chlorophenoxy)-hexyl)-oxirane-2-carboxylic acid ethyl
ester. Examples of other oxirane carboxylic acid compounds useful
in the invention are
2-(4-(3-chlorophenoxy)-butyl)-oxirane-2-carboxylic acid ethyl
ester, 2-(4-(3-trifluoromethylphenoxy)-butyl)-oxirane-2-carboxylic
acid ethyl ester,
2-(5-(4-chlorophenoxy)-pentyl)-oxirane-2-carboxylic acid ethyl
ester, 2-(6-(3,4-dichlorophenoxy)-hexyl)-oxirane-2-carboxylic acid
ethyl ester, 2-(6-(4-fluorophenoxy)-hexyl)-oxirane-2-carboxylic
acid ethyl ester, and 2-(6-phenoxyhexyl)-oxirane-2-carboxylic acid
ethyl ester, the corresponding oxirane carboxylic acids, and their
pharmacologically acceptable salts.
[0034] The foregoing class of oxirane carboxylic acid compounds,
including etomoxir, has been described by Horst Wolf and Klaus
Eistetter in U.S. Pat. No. 4,946,866 for the prevention and
treatment of illnesses associated with increased cholesterol and/or
triglyceride concentration, and by Horst Wolf in U.S. Pat. No.
5,739,159 for treating heart insufficiency. The preparation of
oxirane carboxylic acid compounds, and their use for blood glucose
lowering effects as an antidiabetic agent, is described in Jew et
al U.S. Pat. No. 6,013,666. Etomoxir has been described as an
inhibitor of mitochondrial carnitine palmitoyl transferase-I .by
Mannaerts, G. P., L. J. Debeer, J. Thomas, and P. J. De Schepper.
"Mitochondrial and peroxisomal fatty acid oxidation in liver
homogenates and isolated hepatocytes from control and
clofibrate-treated rats," J. Biol. Chem. 254:4585-4595, 1979.
United States Patent Application 20030036199 by Bamdad et al,
entitled:" Diagnostic tumor markers, drug screening for
tumorigenesis inhibition, and compositions and methods for
treatment of cancer," published Feb. 20, 2003, describes treating a
subject having a cancer characterized by the aberrant expression of
MUC1, comprising administering to the subject etomoxir in an amount
effective to reduce tumor growth.
[0035] The foregoing U.S. Pat. Nos. 4,946,866, 5,739,159, and
6,013,666, United States Patent Application 20030036199, and the
foregoing publication by Mannaerts, G. P., L. J. Debeer, J. Thomas,
and P. J. De Schepper, are incorporated herein by reference. In
addition, U.S. patent application Ser. No. 10/272,432, filed Oct.
15, 2002, entitled "Methods for Regulating Co-Stimulatory Molecule
Expression with Reactive Oxygen," by M. K. Newell, et al. is
incorporated herein by reference in its entirety
[0036] Other, non-limiting examples of fatty acid metabolism
inhibitors include fatty acid transporter inhibitors,
beta-oxidation process inhibitors, reductase inhibitors, and/or
isomerase inhibitors within the fatty acid metabolism pathway.
Specific examples of other fatty acid metabolism inhibitors
include, but are not limited to, cerulenin,
5-(tetradecyloxy)-2-furoic acid, oxfenicine, methyl palmoxirate,
metoprolol, amiodarone, perhexiline, aminocarnitine,
hydrazonopropionic acid, 4-bromocrotonic acid, trimetazidine,
ranolazine, hypoglycin, dichloroacetate, methylene cyclopropyl
acetic acid, and beta-hydroxy butyrate. Structural formulas for
these inhibitors are shown in FIG. 1A-1C. As a another example, the
inhibitor may be a non-hydrolyzable analog of carnitine.
[0037] In one embodiment, the fatty acid metabolism inhibitor is a
carboxylic acid. In some cases, the carboxylic acid may have the
structure: ##STR2## where R comprises an organic moiety, as further
described below. In some cases, R may include at least two nitrogen
atoms, or R may include an aromatic moiety (as further described
below), such as a benzene ring, a furan, etc.
[0038] In another embodiment, the fatty acid metabolism inhibitor
has the structure: ##STR3## where each of R.sup.1 and R.sup.2
independently comprises organic moiety. In some instances, either
or both of R.sup.1 and R.sup.2 may independently be an alkyl, such
as a straight-chain alkyl, for instance, methyl, ethyl, propyl,
etc. In certain cases, R2 may have at least 5 carbon atoms, at
least 10 carbon atoms, or at least 15 or more carbon atoms. For
example, in one embodiment, R.sup.2 may be a tetradecyl moiety. In
other cases, R.sup.2 may include an aromatic moiety, for example, a
benzene ring. In still other cases, R.sup.2 may have the structure:
##STR4## where R.sup.3 comprises an organic moiety and Ar.sup.1
comprises an aromatic moiety. R.sup.3 may be a an alkyl, such as a
straight-chain alkyl. In some instances, Ar.sup.1 may be a benzene
ring or a derivative thereof, i.e., having the structure: ##STR5##
wherein each of R.sup.4, R.sup.5, R.sup.6, R.sup.7, and R.sup.8 is
hydrogen, a halogen, an alkyl, an alkoxy, etc.
[0039] In yet another embodiment, the fatty acid metabolism
inhibitor has the structure: ##STR6## where each of R.sup.10,
R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16
independently comprises hydrogen, a halogen, or an organic moiety,
such as an alkyl, an alkoxy, etc. In some cases, R.sup.10 and
R.sup.11 together may define an organic moiety, such as a cyclic
group. For example, the fatty acid metabolism inhibitor may have
the structure: ##STR7## wherein R.sup.17 comprises an organic
moiety, such as an alkyl, an alkoxy, an aromatic moiety, an amide,
etc. An example, of R.sup.17 is: ##STR8## wherein Ar.sup.2
comprises an aromatic moiety, such as a benzene ring or a benzene
derivative, as previously described.
[0040] In still another embodiment, the fatty acid metabolism
inhibitor includes a dominant negative plasma membrane polypeptide.
The end result of the use (e.g., expression) of a dominant negative
polypeptide in a cell may be a reduction in functional enzymes
present within the fatty acid metabolism pathway. One of ordinary
skill in the art can assess the potential for a dominant negative
variant of a protein or enzyme, and use standard mutagenesis
techniques to create one or more dominant negative variant
polypeptides. For example, one of ordinary skill in the art can
modify the sequence of an enzyme coding region by site-specific
mutagenesis, scanning mutagenesis, partial gene deletion or
truncation, and the like. See, e.g., U.S. Pat. No. 5,580,723 and
Sambrook, et al., Molecular Cloning: A Laboratory Manual, Second
Edition, Cold Spring Harbor Laboratory Press, 1989. One of ordinary
skill in the art then can test the population of mutagenized
polypeptides for diminution in a selected and/or for retention of
such activity of the protein or enzyme. Other similar methods for
creating and testing dominant negative variants of a protein will
be apparent to one of ordinary skill in the art.
[0041] In another set of embodiments, the cells may be exposed to
an agent that inhibits the synthesis or production of one or
enzymes within the fatty acid metabolism pathway. Exposure of the
cells to the agent thus inhibits fatty acid metabolism within the
cell. For example, in one embodiment, an antisense oligonucleotide
may be used that selectively binds to regions encoding enzymes
present within the fatty acid metabolism pathway, such as
2,4-dienoyl-CoA reductase or 2,4-dienoyl-CoA isomerase. Antisense
oligonucleotides are discussed in more detail below.
Glucose Metabolism Inhibitor
[0042] Preferred glucose metabolism inhibitors are 2-deoxyglucose
compounds, defined herein as 2-deoxy-D-glucos, and homologs,
analogs, and/or derivatives of 2-deoxy-D-glucose. While the levo
form is not prevalent, and 2-deoxy-D-glucose is preferred, the term
"2-deoxyglucose" is intended to cover inter alia either
2-deoxy-D-glucose and 2-deoxy-L-glucose, or a mixture thereof. In
general glucose metabolism inhibitors can have the formula:
##STR9## wherein: X represents an O or S atom; R.sub.1 represents a
hydrogen atom or a halogen atom; R.sub.2 represents a hydroxyl
group, a halogen atom, a thiol group, or CO--R.sub.6; and R.sub.3,
R.sub.4, and R.sub.5 each represent a hydroxyl group, a halogen
atom, or CO--R.sub.6 wherein R.sub.6 represents an alkyl group of
from 1 to 20 carbon atoms, and wherein at least two of R.sub.3,
R.sub.4, and R.sub.5 are hydroxyl groups. The halogen atom is as
described above with respect to the oxirane carboxylic acid
compounds, and in R.sub.2, R.sub.3, R.sub.4, and R.sub.5. The
halogen atom is preferably F, and R.sub.6 is preferably a
C.sub.3-C.sub.15 alkyl group.
[0043] Examples of 2-deoxyglucose compounds useful in the invention
are: 2-deoxy-D-glucose, 2-deoxy-L-glucose; 2-bromo-D-glucose,
2-fluoro-D-glucose, 2-iodo-D-glucose, 6-fluoro-D-glucose,
6-thio-D-glucose, 7-glucosyl fluoride, 3-fluoro-D-glucose,
4-fluoro-D-glucose, 1-O-propyl ester of 2-deoxy-D-glucose, 1
-O-tridecyl ester of 2-deoxy-D-glucose, 1-O-pentadecyl ester of
2-deoxy-D-glucose, 3 -O-propyl ester of 2-deoxy-D-glucose,
3-O-tridecyl ester of 2-deoxy-D-glucose, 3-O-pentadecyl ester of
2-deoxy-D-glucose, 4-O-propyl ester of 2-deoxy-D-glucose,
4-O-tridecyl ester of 2-deoxy-D-glucose, 4-O -pentadecyl ester of
2-deoxy-D-glucose, 6-O-propyl ester of 2-deoxy-D-glucose,
6-0-tridecyl ester of 2-deoxy-D-glucose, 6-O-pentadecyl ester of
2-deoxy-D-glucose, and 5-thio-D-glucose, and mixtures thereof.
[0044] A preferred glucose metabolism inhibitor is
2-deoxy-D-glucose, which has the structure: ##STR10## Bifunctional
Compounds
[0045] The bifunctional compounds in their most generic form link a
moiety functioning as a fatty acid metabolism inhibitor to a moiety
functioning as a glycolysis inhibitor. Referring to FIG. 2, generic
forms of five bifunctional compounds, respectively compounds (I) to
(V), are shown, and respective specific preferred bifunctional
compounds are shown in FIG. 3 as respective compounds (VI) to (X),
each of which will be referred to in more detail in the following
examples, each of which link a moiety having the functionality of
an oxirane carboxylic acid to a moiety having the functionality of
a 2-deoxyglucose compound.
EXAMPLE 1
[0046] One can use in the invention, the bifunctional compound (I)
of FIG. 2, having the structure: ##STR11## where R.sub.1 represents
a hydroxyl group, a halogen atom, a thiol group, or CO--R.sub.9
where R.sub.9 represents an alkyl group of from 1 to 20 carbon
atoms, R.sub.4 represents a hydrogen atom or a halogen atom,
R.sub.2, R.sub.3 and R.sub.5 each represent a hydroxyl group, a
halogen atom, or CO--R.sub.9 and where at least two of R.sub.2,
R.sub.3 and R.sub.5 are hydroxyl groups, R.sub.6 and R.sub.7 each
represent a hydrogen atom, a halogen atom, a 1-4 carbon atom alkyl
group, a 1-4 carbon atom alkoxy group, a nitro group or a
trifluoromethyl group, R.sub.8 represents a hydrogen atom or a 1-4
carbon atom alkyl group, X represents O or S, Y represents
(CH.sub.2).sub.k where k is from 2 to 8, or the grouping
--O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1 to 4, n is
a whole number from 2 to 8 wherein the sum of m and n is a whole
number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4.
[0047] A preferred specific bifunctional compound (I) is shown in
FIG. 3 as compound (VI), having the structure: ##STR12##
EXAMPLE 2
[0048] One can use in the invention, the bifunctional compound (II)
of FIG. 2, having the structure: ##STR13## where R.sub.4 represents
a hydrogen atom or a halogen atom, R.sub.2, R.sub.3 and R.sub.5
each represent a hydroxyl group, a halogen atom, or CO--R.sub.9
where R.sub.9 represents an alkyl group of from 1 to 20 carbon
atoms, and where at least two of R.sub.2, R.sub.3 and R.sub.5 are
hydroxyl groups, R.sub.8 represents a hydrogen atom or a 1-4 carbon
atom alkyl group, X represents O or S, and Y represents
(CH.sub.2).sub.k where k is from 2 to 8, or the grouping --O
--(CH.sub.2).sub.m--, m is 0 or a whole number from 1 to 4, n is a
whole number from 2 to 8 wherein the sum of m and n is a whole
number from 2 to 8.
[0049] A preferred specific bifunctional compound (II) is shown in
FIG. 3 as compound (VII), having the structure: ##STR14##
EXAMPLE 3
[0050] One can use in the invention, the bifunctional compound
(III) of FIG. 2, having the structure: ##STR15## where R.sub.1
represents a hydroxyl group, a halogen atom, a thiol group, or
CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to 20
carbon atoms, R.sub.4 represents a hydrogen atom or a halogen atom,
R.sub.2, R.sub.3 and R.sub.5 each represent a hydroxyl group, a
halogen atom, or CO--R.sub.9 and where at least two of R.sub.2,
R.sub.3 and R.sub.5 are hydroxyl groups, R.sub.6 and R.sub.7 each
represent a hydrogen atom, a halogen atom, a 1-4 carbon atom alkyl
group, a 1-4 carbon atom alkoxy group, a nitro group or a
trifluoromethyl group, R.sub.8 represents a hydrogen atom or a 1-4
carbon atom alkyl group, X represents O or S, and Y represents
(CH.sub.2).sub.k where k is from 2 to 8, or the grouping
--O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1 to 4, n is
a whole number from 2 to 8 wherein the sum of m and n is a whole
number from 2 to 8.
[0051] A preferred specific bifunctional compound (III) is shown in
FIG. 3 as compound (VIII), having the structure: ##STR16##
EXAMPLE 4
[0052] One can use in the invention, the bifunctional compound (IV)
of FIG. 2, having the structure: ##STR17## where R.sub.1 represents
a hydroxyl group, a halogen atom, a thiol group, or CO--R.sub.9
where R.sub.9 represents an alkyl group of from 1 to 20 carbon
atoms, R.sub.4 represents a hydrogen atom or a halogen atom,
R.sub.2, R.sub.3 and R.sub.5 each represent a hydroxyl group, a
halogen atom, or CO--R.sub.9 and where at least two of R.sub.2,
R.sub.3 and R.sub.5 are hydroxyl groups, R.sub.6 and R.sub.7 each
represent a hydrogen atom, a halogen atom, a 1-4 carbon atom alkyl
group, a 1-4 carbon atom alkoxy group, a nitro group or a
trifluoromethyl group, R.sub.8 represents a hydrogen atom or a 1-4
carbon atom alkyl group, X represents O or S, Y represents
(CH.sub.2).sub.k where k is from 2 to 8, or the grouping
--O--(CH.sub.2).sub.m--, m is 0 or a whole number from 1 to 4, n is
a whole number from 2 to 8 wherein the sum of m and n is a whole
number from 2 to 8, and Z represents O, S or the grouping
(CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4.
[0053] A preferred specific bifunctional compound (IV) is shown in
FIG. 3 as compound (IX), having the structure: ##STR18##
EXAMPLE 5
[0054] One can use in the invention, the bifunctional compound (V)
of FIG. 2, having the structure: ##STR19## where R.sub.1 represents
a hydroxyl group, a halogen atom, a thiol group, or CO--R.sub.9
where R.sub.9 represents an alkyl group of from 1 to 20 carbon
atoms, R.sub.4 represents a hydrogen atom or a halogen atom,
R.sub.2, R.sub.3 and R.sub.5 each represent a hydroxyl group, a
halogen atom, or CO--R.sub.9 and where at least two of R.sub.2,
R.sub.3 and R.sub.5 are hydroxyl groups, R.sub.6 and R.sub.7 each
represent a hydrogen atom, a halogen atom, a 1-4 carbon atom alkyl
group, a 1-4 carbon atom alkoxy group, a nitro group or a
trifluoromethyl group, R.sub.8 represents a hydrogen atom or a 1-4
carbon atom alkyl group, X represents O or S, and Y represents
(CH.sub.2).sub.k where k is from 2 to 8, or the grouping
--O--(CH.sub.2).sub.m--, where m is 0 or a whole number from 1 to
4, n is a whole number from 2 to 8 wherein the sum of m and n is a
whole number from 2 to 8.
[0055] A preferred specific bifunctional compound (V) is shown in
FIG. 3 as compound (X), having the structure: ##STR20##
[0056] Preferably, the invention provides bifunctional compounds
that link a moiety having the functionality of an oxirane
carboxylic acid compound to a moiety having the functionality of
hypoglycin or a hypoglycin A derivative. Most preferred are
bifunctional compounds that link a moiety having the functionality
of etomoxir to a moiety having the functionality of hypoglycin A.
Referring to FIGS. 4-7, generic forms of twelve bifunctional
compounds (I) to (XII), are shown, and respective specific
preferred bifunctional compounds (XIII) to (XXVII) are shown in
FIGS. 8-12. Each will be referred to in more detail in the
following examples, where R.sub.1, R.sub.2, R.sub.3, R.sub.4, and
R.sub.5 represents a hydroxyl group, a halogen atom, a thiol group,
or CO--R.sub.9 where R.sub.9 represents an alkyl group of from 1 to
20 carbon atoms, R.sub.6 and R.sub.7 each represent a hydrogen
atom, a halogen atom, a 1-4 carbon atom alkyl group, a 1-4 carbon
atom alkoxy group, a nitro group or a trifluoromethyl group,
R.sub.8 represents a hydrogen atom or a 1-4 carbon atom alkyl
group, Y represents (CH.sub.2).sub.k where k is from 2 to 8, 7 or
the grouping --O--(CH.sub.2).sub.m--, m is 0 or a whole number from
1 to 4, n is a whole number from 2 to 8 wherein the sum of m and n
is a whole number from 2 to 8, and Z represents O, S or the
grouping (CH.sub.2).sub.p--O--(CH.sub.2).sub.q or
(CH.sub.2).sub.p--S--(CH.sub.2).sub.q, and p and q are each 0 or a
whole number from 1 to 4.
EXAMPLE 6
[0057] One can use in the invention, the bifunctional compound (I)
of FIG. 4, having the structure: ##STR21##
[0058] A preferred specific example of bifunctional compound (I) is
shown in FIG. 8 as compound (XIII), having the structure:
##STR22##
EXAMPLE 7
[0059] One can use in the invention, the bifunctional compound (II)
of FIG. 4, having the structure: ##STR23##
[0060] A preferred specific example of bifunctional compound (II)
is shown in FIG. 8 as compound (XIV), having the structure:
##STR24##
EXAMPLE 8
[0061] One can use in the invention, the bifunctional compound
(III) of FIG. 5, having the structure: ##STR25##
[0062] A preferred specific example of bifunctional compound (III)
is shown in FIG. 8 as compound (XV), having the structure:
##STR26##
EXAMPLE 9
[0063] One can use in the invention, the bifunctional compound (IV)
of FIG. 5, having the structure: ##STR27##
[0064] A preferred specific example of bifunctional compound (IV)
is shown in FIG. 9 as compound (XVI), having the structure:
##STR28##
EXAMPLE 10
[0065] One can use in the invention, the bifunctional compound (V)
of FIG. 5, having the structure: ##STR29##
[0066] A preferred specific example of bifunctional compound (V) is
shown in FIG. 9 as compound (XVII), having the structure:
##STR30##
EXAMPLE 11
[0067] One can use in the invention, the bifunctional compound (VI)
of FIG. 5, having the structure: ##STR31##
[0068] A preferred specific example of bifunctional compound (VI)
is shown in FIG. 9 as compound (XVIII), having the structure:
##STR32##
EXAMPLE 12
[0069] One can use in the invention, the bifunctional compound
(VII) of FIG. 6, having the structure: ##STR33##
[0070] A preferred specific example of bifunctional compound (VII)
is shown in FIG. 10 as compound (XIX), having the structure:
##STR34##
EXAMPLE 13
[0071] One can use in the invention, the bifunctional compound
(VIII) of FIG. 6, having the structure: ##STR35##
[0072] A preferred specific example of bifunctional compound (VIII)
is shown in FIG. 10 as compound (XX), having the structure:
##STR36##
EXAMPLE 14
[0073] One can use in the invention, the bifunctional compound (IX)
of FIG. 6, having the structure: ##STR37##
[0074] A preferred specific example of bifunctional compound (XXI)
is shown in FIG. 9 as compound (XVII), having the structure:
##STR38##
EXAMPLE 15
[0075] One can use in the invention, the bifunctional compound (X)
of FIG. 7, having the structure: ##STR39##
[0076] A preferred specific example of bifunctional compound (X) is
shown in FIG. 11 as compound (XXII), having the structure:
##STR40##
[0077] Another preferred specific example of bifunctional compound
(X) is shown in FIG. 11 as compound (XXIII), having the structure:
##STR41##
EXAMPLE 16
[0078] One can use in the invention, the bifunctional compound (XI)
of FIG. 7, having the structure: ##STR42##
[0079] A preferred specific example of bifunctional compound (XI)
is shown in FIG. 11 as compound (XXIV), having the structure:
##STR43##
[0080] Another preferred specific example of bifunctional compound
(XI) is shown in FIG. 12 as compound (XXV), having the structure:
##STR44##
EXAMPLE 17
[0081] One can use in the invention, to treat MDR tumors, the
bifunctional compound (XII) of FIG. 7, having the structure:
##STR45##
[0082] A preferred specific example of bifunctional compound (XII)
is shown in FIG. 12 as compound (XXVI), having the structure:
##STR46##
[0083] Another preferred specific example of bifunctional compound
(XII) is shown in FIG. 12 as compound (XXVII), having the
structure: ##STR47##
* * * * *